Bio-3D printing of scaffold-free osteogenic and chondrogenic constructs using rat adipose-derived stromal cells

Abstract

Background: Although autogenous bone implantation is considered to be the gold standard for the reconstruction of bone defects, this approach remains challenging when treating extensive bone defects (EBDs). Therefore, artificial materials (AMs) such as artificial bone and scaffolds are often used for treating EBDs. Nevertheless, complications such as material failure, foreign body reaction, and infection are common. To overcome these issues, we aimed to develop a new treatment for an EBD using scaffold-free adipose-derived stromal cells (ADSCs) to fabricate chondrogenic/osteogenic-induced constructs without AMs. Methods: ADSCs were obtained from the subcutaneous adipose tissue of 8-week-old female Wistar rats (n = 3) and assessed to determine their potential for multilineage differentiation into adipocytes (Oil Red O staining), chondrocytes (hematoxylin and eosin, Alcian blue, and Safranin O staining), and osteoblasts (Alizarin red and von Kossa staining). Spheroids (n = 320), each containing 3.0 × 10 4 ADSCs, were then used to fabricate scaffold-free cell constructs using a bio-3D printer with a needle array. The spheroids and constructs were stimulated with induction medium to induce chondrogenic and osteogenic differentiation. The induced cartilage-and bone-like constructs were finally evaluated using micro-computed tomography (µCT) and histological analysis. Results: The collected ADSCs were capable of trilineage differentiation, and were successfully used to produce scaffold-free constructs. The fabricated constructs (n = 3) exhibited equivalent strength (load, 195.3 ± 6.1 mN; strength, 39.1 ± 1.2 kPa; and stiffness, 0.09 ± 0.01 N/mm) to that of soft tissues such as the muscles in the uninduced condition. In chondrogenic induction experiments, Alcian blue and Safranin O staining confirmed the differentiation of the constructs into cartilage, and cartilage tissue-like structures were produced. In the osteogenic induction experiment, Alizarin Red and von Kossa staining showed calcium salt deposition, and µCT images confirmed the same calcification level as that of the cortical bone. Conclusions: Scaffold-free constructs consisting of ADSCs without an AM were fabricated, and cartilage-and bone-like tissues were successfully generated, demonstrating their potential for bone reconstruction.

Exosomes derived from human adipose‐derived stem cells alleviate hepatic ischemia–reperfusion (I/R) injury through the miR‐183/ALOX5 axis

“…Human ADSCs were obtained from Cellular Engineering Technologies, and its induction was consistent with previous research 32 . HL‐7702 cells were purchased from Shanghai JinYuan Organism (http://www.ssrcc.com.cn/product/?id=181).…”

“…Human ADSCs were obtained from Cellular Engineering Technologies, and its induction was consistent with previous research. 32 HL-7702 cells were purchased from Shanghai JinYuan Organism (http://www.ssrcc.com.cn/ produ ct/?id=181). ADSCs and HL-7702 cells were cultured in 10% fetal bovine serum (FBS)-containing DMEM at 37°C and 5% CO 2 .…”

“…The chondrogenic potential of ADSCs was observed by Alcian blue staining (Figure 1E). In contrast to previous study 32 and the dyeing results HL-7702 cells, we also successfully identified ADSCs based on their specific characteristics.…”

Scaffold-free human vascular calcification model using a bio-three-dimensional printer

“…We fabricated various scaffold-free structures on a bio-3D printer using a method developed in a previous study [15][16][17][18][19][20]. Our group also conducted clinical studies to implant a shunt graft [21,22] in patients undergoing hemodialysis therapy in whom an arteriovenous shunt could not be used because of shunt occlusion, stenosis, or aneurysm.…”